The origins of the circumventricular organs.

The circumventricular organs (CVOs) are specialised neuroepithelial structures found in the midline of the brain, grouped around the third and fourth ventricles. They mediate the communication between the brain and the periphery by performing sensory and secretory roles, facilitated by increased vascularisation and the absence of a blood-brain barrier. Surprisingly little is known about the origins of the CVOs (both developmental and evolutionary), but their functional and organisational similarities raise the question of the extent of their relationship. Here, I review our current knowledge of the embryonic development of the seven major CVOs (area postrema, median eminence, neurohypophysis, organum vasculosum of the lamina terminalis, pineal organ, subcommissural organ, subfornical organ) in embryos of different vertebrate species. Although there are conspicuous similarities between subsets of CVOs, no unifying feature characteristic of their development has been identified. Cross-species comparisons suggest that CVOs also display a high degree of evolutionary flexibility. Thus, the term 'CVO' is merely a functional definition, and features shared by multiple CVOs may be the result of homoplasy rather than ontogenetic or phylogenetic relationships.

Direct and indirect roles of Fgf3 and Fgf10 in innervation and vascularisation of the vertebrate hypothalamic neurohypophysis.

The neurohypophysis is a crucial component of the hypothalamo-pituitary axis, serving as the site of release of hypothalamic neurohormones into a plexus of hypophyseal capillaries. The growth of hypothalamic axons and capillaries to the forming neurohypophysis in embryogenesis is therefore crucial to future adult homeostasis. Using ex vivo analyses in chick and in vivo analyses in mutant and transgenic zebrafish, we show that Fgf10 and Fgf3 secreted from the forming neurohypophysis exert direct guidance effects on hypothalamic neurosecretory axons. Simultaneously, they promote hypophyseal vascularisation, exerting early direct effects on endothelial cells that are subsequently complemented by indirect effects. Together, our studies suggest a model for the integrated neurohemal wiring of the hypothalamo-neurohypophyseal axis.

FGF-dependent midline-derived progenitor cells in hypothalamic infundibular development.

The infundibulum links the nervous and endocrine systems, serving as a crucial integrating centre for body homeostasis. Here we describe that the chick infundibulum derives from two subsets of anterior ventral midline cells. One set remains at the ventral midline and forms the posterior-ventral infundibulum. A second set migrates laterally, forming a collar around the midline. We show that collar cells are composed of Fgf3(+) SOX3(+) proliferating progenitors, the induction of which is SHH dependent, but the maintenance of which requires FGF signalling. Collar cells proliferate late into embryogenesis, can generate neurospheres that passage extensively, and differentiate to distinct fates, including hypothalamic neuronal fates and Fgf10(+) anterior-dorsal infundibular cells. Together, our study shows that a subset of anterior floor plate-like cells gives rise to Fgf3(+) SOX3(+) progenitor cells, demonstrates a dual origin of infundibular cells and reveals a crucial role for FGF signalling in governing extended infundibular growth.

Pituitary melanotrope cells of Xenopus laevis are of neural ridge origin and do not require induction by the infundibulum.

Classical studies in amphibians have concluded that the endocrine pituitary and pars intermedia are derived from epithelial buccal epidermis and do not require the infundibulum for their induction. These studies also assumed that the pituitary is not subsequently determined by infundibular induction. Our extirpation, auto-transplantation and immunohistochemical studies with Xenopus laevis were initiated to investigate early presumptive pituitary development. These studies were conducted especially with reference to the pars intermedia melanotrope cell's induction, and its production and release of α-melanophore stimulating hormone (α-MSH) from the precursor protein proopiomelanocortin (POMC). Auto-transplantation studies demonstrated that the pituitary POMC-producing cells are determined at a stage prior to pituitary-infundibular contact. The results of experiments involving the extirpation of the presumptive infundibulum also indicated that the infundibulum is not essential for the differentiation of POMC-producing cells. We also demonstrated that early pituitary development involves adherence to the prechiasmatic area of the diencephalon with the pituitary placode growing in a posterior direction toward the infundibulum where contact occurs at Xenopus stage 39/40. Overall, our studies provide a model for early tissue relations among presumptive pituitary, suprachiasmatic nucleus, pars tuberalis and infundibulum during neurulation and later neural tube stages of development. It is hypothesized that the overlying chiasmatic area suppresses pituitary differentiation.

A role of the LIM-homeobox gene Lhx2 in the regulation of pituitary development.

The mammalian pituitary gland originates from two separate germinal tissues during embryonic development. The anterior and intermediate lobes of the pituitary are derived from Rathke's pouch, a pocket formed by an invagination of the oral ectoderm. The posterior lobe is derived from the infundibulum, which is formed by evagination of the neuroectoderm in the ventral diencephalon. Previous studies have shown that development of Rathke's pouch and the generation of distinct populations of hormone-producing endocrine cell lineages in the anterior/intermediate pituitary lobes is regulated by a number of transcription factors expressed in the pouch and by inductive signals from the ventral diencephalon/infundibulum. However, little is known about factors that regulate the development of the posterior pituitary lobe. In this study, we show that the LIM-homeobox gene Lhx2 is extensively expressed in the developing ventral diencephalon, including the infundibulum and the posterior lobe of the pituitary. Deletion of Lhx2 gene results in persistent cell proliferation, a complete failure of evagination of the neuroectoderm in the ventral diencephalon, and defects in the formation of the distinct morphological features of the infundibulum and the posterior pituitary lobe. Rathke's pouch is formed and endocrine cell lineages are generated in the anterior/intermediate pituitary lobes of the Lhx2 mutant. However, the shape and organization of the pouch and the anterior/intermediate pituitary lobes are severely altered due to the defects in development of the infundibulum and the posterior lobe. Our study thus reveals an essential role for Lhx2 in the regulation of posterior pituitary development and suggests a mechanism whereby development of the posterior lobe may affect the development of the anterior and intermediate lobes of the pituitary gland.

Embryonic morphogenesis of the human pituitary.

Light microscopy methods were used to study the main stages in the organogenesis of the pituitary in human embryos at Carnegie stages (CS) 12-23. Rathke's pouch (RP) was shown to form as a traction fold over whole width of the roof of the stomodeum in embryos at CS 12 due to a flexure of the neural tube with which the epithelium had a tight anatomical relationship (the attached part of the anterior wall of the RP) in the median plane of the embryo. The rudiment of the hypothalamic infundibulum and neurohypophysis formed at CS 15, as a thickening of the posterior wall of the diencephalon. Transorientation of the positions of brain components, including the rudiment of the hypophysis, occurred at CS 20-23. The attached part of the anterior wall of the RP then formed the pars intermedia and pars tuberalis of the anterior lobe, while the epithelium of the orifice of the RP and its posterior wall formed the pars distalis. From CS 20 to 23, the RP epithelium formed the structural-functional units of the adenohypophysis, i.e., the epithelial cords, by invagination.

The unique endocrine milieu of the fetus.

Table II summarizes in tabular form the major features of the fetal endocrine milieu discussed in the foregoing pages. The mammalian fetus develops in an environment where respiration, alimentation, and excretory functions are provided by the placenta. Fetal tissue metabolism is oriented largely to anabolism; body temperature is modulated by maternal metabolism, and fetal tissue thermogenesis is maintained at a basal level. Tissue and organ growth appear to be regulated by growth factors which probably function by autocrine or paracrine mechanisms during most of gestation (72, 146-148). In this milieu conventional endocrine control systems are largely redundant, and other transient systems more appropriate to the intrauterine environment have evolved. We have developed some insights into these systems, but much more information is necessary before we can truly understand this fascinating environment.

Micromolar 4-aminopyridine enhances invasion of a vertebrate neurosecretory terminal arborization: optical recording of action potential propagation using an ultrafast photodiode-MOSFET camera and a photodiode array.

Modulation of the amount of neuropeptide released from a neurosecretory tissue may be achieved by different means. These include alterations in the quantity secreted from each active nerve terminal or in the actual number of terminals activated. From the vertebrate hypothalamus, magnocellular neurons project their axons as bundles of fibers through the median eminence and infundibular stalk to arborize extensively and terminate in the neurohypophysis, where the neurohypophysial peptides and proteins are released into the circulation by a Ca-dependent mechanism. Elevating [Ca2+]o increases the magnitude of an intrinsic optical change in the neurohypophysial terminals that is intimately related to the quantity of neuropeptide released. Similarly, the addition of micromolar concentrations of 4-aminopyridine to the bathing solution enhances this change in large angle light scattering. However, we show here that, while these effects are superficially similar, they reflect different mechanisms of action. Evidence from intrinsic optical signals (light scattering) and extrinsic (potentiometric dye) absorption changes suggests that calcium increases the amount of neuropeptide released from each active terminal in the classical manner, while 4-aminopyridine exerts its secretagogue action by enhancing the invasion of action potentials into the magno-cellular neuron's terminal arborization, increasing the actual number of terminals activated. Physiologically, electrical invasion of the complex terminal arborization in the neurohypophysis may represent an extremely sensitive control point for modulation of peptide secretion. This would be especially effective in a neurohaemal organ like the posterior pituitary, where, in contrast with a collection of presynaptic terminals, the precise location of release is less important than the quantity released.

Adrenocorticotropin secretion by fetal sheep anterior and intermediate lobe pituitary cells in vitro: effects of gestation and adrenalectomy.

Immunoreactive (ir) ACTH is present in the fetal sheep intermediate lobe (IL) as well as the anterior pituitary (AP). It is not clear whether fetal IL cells can secrete irACTH and if gestational age and glucocorticoids influence the secretion of ACTH from these tissues in a similar fashion. Therefore, we examined the control of irACTH secretion by IL cells, whether the responsiveness of AP and IL cells to arginine vasopressin (AVP) and CRH changes during gestation, and whether withdrawal of adrenal steroids by adrenalectomy influences AP and IL responses. Cultured pituitary cells were studied from intact fetuses at an immature (n = 5; 108 +/- 5 days) and a mature (n = 8; 139 +/- 0 days) stage, from mature fetuses 3 weeks after bilateral adrenalectomy (n = 6), and from neonatal lambs within 16 h of birth (n = 6). Secretion of irACTH was determined by RIA of incubation medium obtained during 3-h exposure of cells to vehicle, AVP, CRH, or both. In all cases, IL cells secreted measurable irACTH. The IL cells of immature fetuses responded to CRH (133 +/- 8% increase over basal secretion), AVP (52 +/- 6%), and CRH plus AVP (244 +/- 8%). In contrast, IL cells from mature fetuses responded only to CRH (160 +/- 20%) or CRH plus AVP (259 +/- 44%), as did cells from mature adrenalectomized fetuses (CRH, 356 +/- 70%; CRH plus AVP, 627 +/- 100%). Secretion from neonatal IL cells was not significantly increased above basal rates by CRH and/or AVP. The AP cells from immature fetuses responded significantly to CRH (406 +/- 16%), AVP (114 +/- 8%), and CRH plus AVP (559 +/- 38%), whereas cells from mature fetuses responded only to AVP (249 +/- 40%) or to CRH plus AVP (570 +/- 146%). In AP cells from mature adrenalectomized fetuses, the response pattern resembled that of immature intact fetal sheep (CRH, 429 +/- 76%; AVP, 146 +/- 15%; CRH plus AVP, 541 +/- 94%). Neonatal AP cells responded to CRH (196 +/- 25%), AVP (442 +/- 71%), and CRH plus AVP (646 +/- 93%). Further characterization of IL cells (n = 6 fetal and 2 neonatal) indicated that they were inhibited by dopamine (basal ACTH secretion decreased by 25 +/- 4%; ACTH secretory response to CRH decreased by 32 +/- 10%). These results show that fetal neurointermediate lobe cells secrete irACTH under basal and stimulated conditions. Moreover, the pattern of response of AP and neurointermediate lobe cells to secretagogues is influenced by gestational age and, possibly, cortisol.

Neurohypophysial hormones of the 1-month-old bovine fetus: absence of vasotocin during mammal development.

The neurohypophysial hormones of the 1-month-old bovine fetus have been identified by their positions in ion-exchange chromatography and their retention times in high-pressure reverse-phase partition chromatography. Arginine vasopressin and oxytocin have been recognized. The molar ratio vasopressin/oxytocin in neurohypophysis is about 6 in the 1-month-old fetus compared with 4 in the 3-month-old fetus, 2.7 in the 7-month-old fetus and 1 in the adult. Vasotocin is virtually absent even in the early fetus (less than 0.1% of arginine vasopressin). The occurrence of a vasotocin gene expressed in the fetus but silent in the adult appears unlikely.

Projections from the hypothalamus to the posterior lobe in rats during ontogenesis: 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate tracing study.

The objective of this study was to determine the schedule of the arrival of the axons from the hypothalamus to the posterior lobe of the pituitary (PL) in rats during ontogenesis by using the fluorescent lipophilic carbocyanine dye 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI) as a retrograde tracer. After preliminary fixation of the brain, DiI crystals were implanted in the PL on embryonic day 15 (E15), E16, E17, and E19 as well as on postnatal day 2 (P2) and P9. This was followed by a DiI retrograde diffusion along the plasma membrane and subsequent staining of hypothalamic neuronal cell bodies. The supraoptic nucleus (SO) contained an accumulation of fluorescent cells that extended toward the diamond-like swelling of the third ventricle as early as E15. These data suggest that the magnocellular neurons of the SO send their axons to the PL at the very beginning of differentiation, perhaps even before reaching their final position. The initial axons of the neurons of the paraventricular nucleus proper (PV) appeared to reach the PL significantly later, at E17. In addition to the SO and the PV, accessory magnocellular nuclei contributed to the innervation of the PL in perinatal rats. The neurons of the retrochiasmatic accessory nucleus first sent their axons to the PL on E16-E17. Axons that originated from other accessory hypothalamic nuclei reached the PL after birth, suggesting a delay in their involvement in the regulation of visceral functions compared with other magnocellular nuclei. Thus, the axons of magnocellular neurons reach the PL unexpectedly early in embryogenesis, raising the possibility of the functional significance of vasopressin and oxytocin as fetal neurohormones.

Lipocortin-1 immunoreactivity in the human pituitary gland.

We evaluated the distribution of lipocortin-1 immunoreactivity in 118 immature or mature human hypophyses using the peroxidase-antiperoxidase (PAP) technique with a polyclonal rabbit antiserum against lipocortin-1. Serial sections were evaluated for five pituitary hormones and S-100 protein immunoreactivity to compare their distributions with that of lipocortin-1. Scattered or moderate numbers of cells exhibited lipocortin-1 immunoreactivity in the pars distalis of 89 subjects ranging in age from 27 weeks' gestation to 83 years. Seven immature and seven aged specimens exhibited no immunostaining, while 15 specimens from older individuals exhibited only rare immunostaining. Immunostaining did not appear to co-localize selectively with any specific pituitary hormone, although the distribution of immunoreactivity did overlap that of some corticotrophs and was seen in elongated processes of S-100-containing folliculostellate cells. Lipocortin-1 was also found in epithelial cells lining colloid cysts of the residual pars intermedia in 115 of 118 pituitaries ranging in age from 23 weeks' gestation to 83 years. In many intermediate lobe cysts, lipocortin-1 exhibited a pattern of immunoreactivity that partially overlapped the distribution of S-100 protein immunostaining, although the pattern was not identical. Pre-absorption of anti-lipocortin-1 antiserum with lipocortin-1-coupled Sepharose-4B immunoreactivity resulted in loss of immunoreactivity in both lobes. No lipocortin-1 immunoreactivity was seen in the neurohypophysis.

Ontogeny of neurophysin in the rat pituitary gland. An electron microscope immunohistochemical study.

The ontogenic development of neurophysin has not been investigated so far. To study this problem, electron microscopic immunohistochemistry was performed on ultrathin sections of the posterior pituitary of foetal and newborn rats. A concomitant appearance of secretory granules and neurophysin was first noted in the posterior pituitary of 18-day foetus. Neurophysin was detected only in the secretory granules, all of which were labelled. The number of granules (75-110 nm in diameter) as well as the intensity of the reaction for neurophysin increased as the foetus became older. In the newborn rat, the secretory granules were more numerous and showed a strong neurophysin positive reaction. The results support the hypothesis that neurophysin is an essential component for the formation of the secretory granules in the hypothalamo-neurophypophysial system.

Anatomical observations of the development of the pituitary capsule.

A morphological study on the development of the pituitary capsule is reported, based on serial or step-sections of 56 pituitary glands from human embryos and fetuses ranging in gestational age from 4 to 40 weeks. Formation of the pituitary capsule is preceded at a very early embryonic stage by mesenchymal cell proliferation around Rathke's pouch when it forms the primitive adenohypophysis. The proportionally large adenohypophysis wraps around the neurohypophysis as it grows down from the diencephalon. Therefore, the fibrous capsule around the adenohypophysis represents the whole hypophysis as a membrane district from the dura mater or pia arachnoid membrane. The pituitary capsule appears not be to a derivative of either the dura mater or the pia arachnoid; instead it is a separate covering of the developing Rathke's pouch.

An electron microscope study of the differentiating capillaries of the mouse neurohypophysis.

The developing capillaries of the mouse neurohypophysis were studied in the electron microscope to elucidate the fine structural differentiation of the vascular component of the neuro-hemal contact zones in the external median eminence and the neural lobe. In the embryo the growth of the superficial net of the primary plexus of the hypophysial portal system is largely manifested by the presence of proliferation areas located within the capillary plexus covering the surface of the median eminence. Presumptive shallow capillary loops diverge from these ares in the first postnatal week. Differentiation of the capillary wall follows the pattern outlined for continuous capillaries. A few fenestrae appear in the endothelium of immature, superficial vessels at the 17th gestational day, increase in frequency during the following embryonic days, and occur regularly in the postnatal animal. In the neural lobe the internal capillaries proliferative by vascular sprouts emanating from the vessels on the surface of the gland. At the end of embryonic time an extensive net has developed, composed of capillaries with immature characteristics. Proliferation is largely finished by the end of the third postnatal week, when mature capillaries dominate the picture. Formation of attenuated, porous areas is a postnatal process, apart from single fenestrae appearing in the walls of a few immature capillaries in late fetal life. The structural possibilities for an onset of neurohypophysial function in the mouse is discussed.

The human brain at stage 16, including the initial evagination of the neurohypophysis.

Thirty-nine sectioned embryos of stage 16 were studied. Up to this stage the amygdaloid body is derived entirely from the medial eminence, which was purely diencephalic in stage 14, but now extends also to the telencephalon. The area of the future olfactory bulb is indicated by the presence of olfactory fibres entering the brain wall; the future olfactory tubercle is characterized by cellular islands. The presence of the hippocampal thickening and various histological features make it possible to outline the main, future cortical areas already at this early stage: archi-, paleo-, and neopallium. Hippocampus and area dentata correspond to the areas identified by Hines (1922) and Bartelmez and Dekaban (1962) but not to those identified by Humphrey (1966). The interventricular foramen is wide. The cerebral hemispheres grow rostrally and dorsally, thereby forming the beginning of the longitudinal fissure. Apart from the commissure of the superior colliculi, which began to appear in advanced embryos of stage 14, fibres of the posterior commissure are now present in some specimens. The neurohypophysis is apparent in fewer than half of the embryos. The marginal ridge (zona limitans intrathalamica) separates the dorsal from the ventral thalamus. Cranial nerve 3 emerges from M2. M1 has become shorter. Important pathways are beginning: the olfactory route by the olfactory fibres and the medial forebrain bundle; the vestibular by vestibulocerebellar and vestibulospinal fibres; gustatory by chorda tympani, nervus intermedius, and tractus solitarius. Fibres of the cochlear nerve are noted. The first parasympathetic ganglia, submandibular and ciliary, are identifiable. Asymmetry of the cerebral hemispheres was noted in one specimen.

Early development of the pituitary gland in Acipenser naccarii (Chondrostei, Acipenseriformes): an immunocytochemical study.

The distribution and appearance of secretory cells in the pituitary gland were investigated for the first time in a chondrostean species, Acipenser naccarii, from embryos to juveniles, by immunohistochemistry with mammalian and teleost hormone antisera. On 5.5 day post-fertilization (2.5 days pre-hatching), the pituitary of embryos appears as an oval cell mass with a narrow central cavity (hypophysial cleft), close to the ventral border of diencephalon under the third ventricle. At that time no neurohypophysis is observed, the adenohypophysis is not yet structurally divided into pars intermedia (PI) and pars distalis (PD) and only immunoreactive growth hormone cells are detectable. Seven days post-fertilization (1 day pre-hatching) the immunoreactive thyrotropic cells appear in the ventral region and the immunoreactive adrenocorticotropic cells in the posterior dorsal one. At hatching, some immunoreactive melanotropic (ir-MSH) cells are visible in the posterior dorsal region and some immunoreactive prolactin cells in the anterior one. Eight days later the immunoreactive somatolactin cells appear along the posterior dorsal border and the immunoreactive gonadotropic I (ir-GtH I) cells in the ventral region. Here, a few ir-GtH II cells finally appear in 76-86 day old juveniles. The gland elongates after hatching and in 8-day-old larvae two adenohypophysial regions are identified: a posterior (the presumptive PI) and an anterior one (the presumptive PD). In 156-166-day-old juveniles three regions (rostral and proximal pars distalis and pars intermedia) appear and a high number of ir-MSH cells are visible in the rostral region. The first protrusion of neurohypophysis into adenohypophysis is observed in 76-86-day-old juveniles and increases with age, branching into PI. The rostro-caudal distribution of the immunoreactive cells follows the spatial expression of the corresponding hormone gene families observed in zebra fish, suggesting similar differentiating mechanisms in teleosts and chondrosteans.

Maternal-fetal neurohypophyseal system.

The author reviews our present understanding of neurohypophyseal peptide metabolism and physiology, an understanding that is largely based upon the isolation and accurate measurement of arginine vasopressin, oxytocin, and arginine vasotocin. Discussed are both the major advances in understanding and the areas of inquiry that remain or that have been newly opened.

Development pharmacokinetics of the posterior pituitary hormones.

Arginine vasotocin, arginine vasopressin, and oxytocin play a critical role in the stimulation of labor and delivery and in salt and water homeostasis in the newborn infant. The authors present information on their chemistry, secretion, and metabolism, and discuss the clinical effects upon target organs of their presence or absence.

Development of the neurogliohemal complex in the mouse neurohypophysis.

The mouse neurohypophysis was studied at different ages of development in order to analyse the ultrastructural changes that lead to the maturation of the neurogliohemal complex and to determine the existence of permeability between the blood capillaries and the neurohypophysial channels. In all the studies ages, two groups of 5 animals each were intravenously injected with different tracer solutions: to one group, 10 microliters of cationized ferritin were used and to the other, 10 microliters of ferrous fumarate were applied. For the ultrastructural studies the tissue samples were processed using the conventional techniques for electron microscopy. At day 17 of prenatal age, some hypothalamic axons (10 axonic profiles/20 microns2) were already seen within the neurohypophysis, increasing threefold (26 to 30 axonic profiles/20 microns2) at prenatal day 19. In these axons terminals, the first neurosecretory vesicles began to appear. At this early age, the glial cells formed few prolongations. Between postnatal days 1 and 9, numerous axon terminals containing dense neurosecretory vesicles composed the neuropile areas. After day 9, there was a broadening of the intercellular space, which we have termed as neurohypophysial channels; these were actually expansions of the existing extracellular space in the neurohypophysis. Between days 9 and 21, the population of axon terminals showing a higher density of neurosecretory vesicles continued to increase in number. Some of these axon terminals were separated by irregular neurohypophysial channels. The glial cells showed scarce cytoplasm and formed numerous lamellar prolongations, which became increasingly finer surrounding bundles of individual axons.(ABSTRACT TRUNCATED AT 250 WORDS)